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Briefly, cells were incubated with equal volume of virus (MOI = 50 for the control virus) for 45 min on ice, and the bound virus was allowed to be internalized at 37 °C for 25 min.
The volume of virus added was determined by previously calculating the 200TCID50 value.
An equal volume of virus, adjusted to approximately 8HAunits/50 microliter was added to each well.
This may be due to the weaker anaesthesia or the lesser volume of virus solution used, leading to less stress in the animals.
For applications requiring sparse labeling, efficiency can be reduced by decreasing the volume of virus injected, and by directing imaging studies to the periphery of viral transfection.
A 2-ul volume of virus (rAAV-XIAP or rAAV-GFP), diluted 50 1 with fluorescein tracer was injected into the sub-retinal space of the eye, in the central retina, nasal to the optic disc.
Similar(33)
For mock infection, cells were exposed to an equal volume of virus-free buffer.
At 2 h postinfection, another volume of virus-free growth medium was added.
The volume of virus-serum inoculum needed per well in 24-well plates was also assessed.
Further, much smaller volumes of virus pools are required for the fingerpad method while allowing for replicates and controls being tested on the same subject in any given sitting.
A glass pipette was used to pressure inject small volumes of virus (20 100 nl per injection site).
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